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目的探讨RNA干扰(RNAi)技术使人X射线修复交叉互补组基因1(XRCC1)沉默后人支气管上皮细胞(HBE)的生物学性状变化。方法研究设实验组(转染重组质粒的HBE)、空载体组(转染空载体的HBE)、正常对照组(正常HBE)。XRCC1干扰后的HBE的生物学性状分析运用载体介导的RNAi技术抑制XRCC1在HBE中的表达;在光学显微镜下观察实验组和正常对照组细胞的形态学特征;对实验组、空载体组及正常对照组绘制细胞生长曲线,分析细胞倍增时间,运用流式细胞技术分析细胞周期;以汉丽埃塔(HeLa)细胞作阳性对照,采用软琼脂糖恶性程度鉴定实验分析实验组细胞的恶性程度。结果实验组细胞体积稍有增大,细胞内颗粒物明显增多;实验组、空载体组及正常对照组细胞的生长曲线均呈现相似的“S”型;实验组、空载体组及正常对照组细胞的倍增时间相近,分别为0.70、0.83、0.74 d;3组细胞的G1期、G2期和S期分布相近;实验组细胞未见在双层软琼脂糖中生长出细胞集落,而阳性对照细胞集落明显。结论 XRCC1基因缺陷对HBE生物学性状的短期影响不明显。
Objective To investigate the biological characteristics of human bronchial epithelial cells (HBE) after RNA interference (RNAi) technology silences human XRCC1 gene. Methods The experimental group (HBE transfected with recombinant plasmids), empty vector group (HBE transfected with empty vector) and normal control group (normal HBE) were established. XRCC1 after the interference of biological characteristics of HBE using vector-mediated RNAi inhibition of XRCC1 expression in HBE; observed under optical microscope morphological characteristics of the experimental group and normal control cells; the experimental group, empty vector group and The normal control group were drawn cell growth curve, cell doubling time was analyzed, the cell cycle was analyzed by flow cytometry; HeLa cells as positive control, the malignant degree . Results The cell volume of the experimental group increased slightly and the intracellular particles increased obviously. The growth curves of the experimental group, the empty vector group and the normal control group showed similar “S” shape; the experimental group, the empty vector group and the normal control group The doubling time of the group of cells was similar (0.70,0.83,0.74 d, respectively). The distribution of G1 phase, G2 phase and S phase were similar in all the three groups. No colonies were found in the double soft agar in the experimental group, but positive Control cell colonies significantly. Conclusion The short-term effect of XRCC1 gene defect on the biological characteristics of HBE is not obvious.